Arango Julián Camilo, Puerta-Arias Juan David, Pino-Tamayo Paula Andrea, Salazar-Peláez Lina María, Rojas Mauricio, González Ángel
Medical and Experimental Mycology Group, Corporación para Investigaciones Biológicas (CIB)-Universidad de Antioquia, Medellín, Colombia.
School of Microbiology, Universidad de Antioquia, Medellín, Colombia.
PLoS Negl Trop Dis. 2017 Oct 17;11(10):e0006006. doi: 10.1371/journal.pntd.0006006. eCollection 2017 Oct.
Bone marrow-derived mesenchymal stem cells (BMMSCs) have been consider as a promising therapy in fibrotic diseases. Experimental models suggest that BMMSCs may be used as an alternative therapy to treat chemical- or physical-induced pulmonary fibrosis. We investigated the anti-fibrotic potential of BMMSCs in an experimental model of lung fibrosis by infection with Paracoccidioides brasiliensis. BMMSCs were isolated and purified from BALB/c mice using standardized methods. BALB/c male mice were inoculated by intranasal infection of 1.5x106 P. brasiliensis yeasts. Then, 1x106 BMMSCs were administered intra venous at 8th week post-infection (p.i.). An additional group of mice was treated with itraconazole (ITC) two weeks before BMMSCs administration. Animals were sacrificed at 12th week p.i. Histopathological examination, fibrocytes counts, soluble collagen and fibrosis-related genes expression in lungs were evaluated. Additionally, human fibroblasts were treated with homogenized lung supernatants (HLS) to determine induction of collagen expression. Histological analysis showed an increase of granulomatous inflammatory areas in BMMSCs-treated mice. A significant increase of fibrocytes count, soluble collagen and collagen-3α1, TGF-β3, MMP-8 and MMP-15 genes expression were also observed in those mice. Interestingly, when combined therapy BMMSCs/ITC was used there is a decrease of TIMP-1 and MMP-13 gene expression in infected mice. Finally, human fibroblasts stimulated with HLS from infected and BMMSCs-transplanted mice showed a higher expression of collagen I. In conclusion, our findings indicate that late infusion of BMMSCs into mice infected with P. brasiliensis does not have any anti-fibrotic effect; possibly because their interaction with the fungus promotes collagen expression and tissue remodeling.
骨髓间充质干细胞（BMMSCs）已被视为纤维化疾病的一种有前景的治疗方法。实验模型表明，BMMSCs可作为治疗化学或物理诱导的肺纤维化的替代疗法。我们通过巴西副球孢子菌感染，在肺纤维化实验模型中研究了BMMSCs的抗纤维化潜力。使用标准化方法从BALB/c小鼠中分离并纯化BMMSCs。通过鼻内感染1.5×10⁶个巴西副球孢子菌酵母对BALB/c雄性小鼠进行接种。然后，在感染后第8周静脉注射1×10⁶个BMMSCs。另一组小鼠在注射BMMSCs前两周用伊曲康唑（ITC）治疗。在感染后第12周处死动物。评估肺组织的组织病理学检查、纤维细胞计数、可溶性胶原蛋白和纤维化相关基因表达。此外，用人成纤维细胞处理肺匀浆上清液（HLS）以确定胶原蛋白表达的诱导情况。组织学分析显示，接受BMMSCs治疗的小鼠肉芽肿性炎症区域增加。在这些小鼠中还观察到纤维细胞计数、可溶性胶原蛋白以及胶原蛋白-3α1、TGF-β3、MMP-8和MMP-15基因表达显著增加。有趣的是，当使用BMMSCs/ITC联合治疗时，感染小鼠中TIMP-1和MMP-13基因表达降低。最后，用感染和移植BMMSCs小鼠的HLS刺激人成纤维细胞，显示I型胶原蛋白表达更高。总之，我们的研究结果表明，在感染巴西副球孢子菌的小鼠中晚期注入BMMSCs没有任何抗纤维化作用；可能是因为它们与真菌的相互作用促进了胶原蛋白表达和组织重塑。
